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. 2009 Jul 15;15(9-10):297–306. doi: 10.2119/molmed.2009.00030

Figure 6. Expression levels of different cell cycle regulators following E2F1, cyclin E1/E2 depletionand effect of the combined administration of different siRNAs on BrdU incorporation.

Figure 6

A) Three days after transfection, the protein levels of different cell cycle regulators were measured in protein extracts obtained from CSMCs of donor 2: shown are representative blots; # all the numbers reported below the blots derive from band quantification normalized to GAPDH and are expressed as ratio of siGL2 treatment; for ppRb (hyper-phosphorylated pRB) and cyclin A2, whose phosphorylation status and protein levels, respectively, were modified, cumulative data are reported as ratio to GAPDH; data are expressed as means ± SEM, n=3, + p< 0.05 compared to control; B) mRNA levels of cyclin A2 and Cdk2 were measured in CSMC treated with siE2F1-1324 in total RNA extracts obtained from both donors (28S transcript levels served as normalizator); results are expressed as means ± SEM, n=6, * p< 0.05 compared to control. C) CSMCs were treated by a mix of the different siRNAs (115 or 76 nM each in the case of a combination of two or three siRNA, respectively) and CSMC proliferation evaluated by BrdU incorporation; the effects of the combined administration were compared to an equimolar concentration of each of the single siRNAs (230 nM each) three days after transfection; NTC = non-treated cells, siGL2 = control siRNA treated cells; cumulative data from donor 1 and 2 CSMCs are expressed as means ± SEM, n=3.